Anti-USP48 antibody [EPR26175-11] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-USP48 antibody [EPR26175-11] - BSA and Azide free (AB314544)

This data was developed using ab314543, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (human embryonic kidney epithelial cell) cells labelling USP48 with ab314543 at 1/500 dilution (0.1 ug, Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-USP48 antibody [EPR26175-11] - BSA and Azide free (AB314544)

This data was developed using ab314543, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling USP48 with ab314543 at 1/50 (10.44 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2ug/ml) dilution (Green). Confocal image showing nuclear staining in 293T cells. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2ug/ml) dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-USP48 antibody [EPR26175-11] - BSA and Azide free (AB314544)

This data was developed using ab314543, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 80% Methanol-fixed, 0.1% TritonX-100 permeabilized K-562 (human chronic myelogenous leukemia lymphoblast) cells labelling USP48 with ab314543 at 1/50 (10.44 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1001 (2ug/ml) dilution (Green). Confocal image showing nuclear staining in K-562 cells. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1001 (2ug/ml) dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-USP48 antibody [EPR26175-11] - BSA and Azide free (AB314544)

This data was developed using ab314543, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized K-562 (human chronic myelogenous leukemia lymphoblast) cells labelling USP48 with ab314543 at 1/500 dilution (0.1 ug, Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP48 antibody [EPR26175-11] - BSA and Azide free (AB314544)

This data was developed using ab314543, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human gastric carcinoma tissue labeling USP48 with ab314543 at 1/500 (1.044 ug/ml) followed by a ready to use rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human gastric carcinoma. The section was incubated with ab314543 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP48 antibody [EPR26175-11] - BSA and Azide free (AB314544)

This data was developed using ab314543, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human lung tissue labeling USP48 with ab314543 at 1/500 (1.044 ug/ml) followed by a ready to use rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human lung. The section was incubated with ab314543 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP48 antibody [EPR26175-11] - BSA and Azide free (AB314544)

This data was developed using ab314543, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded rat lung tissue labeling USP48 with ab314543 at 1/1000 (0.522 ug/ml) followed by a ready to use rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat lung. The section was incubated with ab314543 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP48 antibody [EPR26175-11] - BSA and Azide free (AB314544)

This data was developed using ab314543, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling USP48 with ab314543 at 1/1000 (0.522 ug/ml) followed by a ready to use rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse lung. The section was incubated with ab314543 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

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Western blot - Anti-USP48 antibody [EPR26175-11] - BSA and Azide free (AB314544)

This data was developed using ab314543, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST The molecular weight observed is consistent with what has been described in the literature (PMID : 28233861; PMID : 34445214). In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-USP48 antibody [EPR26175-11] (<a href='/en-us/products/primary-antibodies/usp48-antibody-epr26175-11-ab314543'>ab314543</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

293T transfected with siRNA specifically targeti USP48 whole cell lysate at 20 µg

Lane 3:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 4:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 145 kDa,160 kDa

false

Exposure time: 92s

• WB

Supplier Data

Western blot - Anti-USP48 antibody [EPR26175-11] - BSA and Azide free (AB314544)

This data was developed using ab314543, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-USP48 antibody [EPR26175-11] (<a href='/en-us/products/primary-antibodies/usp48-antibody-epr26175-11-ab314543'>ab314543</a>) at 1/1000 dilution

Lane 1:

Human hippocampus tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Rat brain tissue lysate at 20 µg

Lane 4:

Rat lung tissue lysate at 20 µg

Secondary

Lanes 1 - 4:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/2000 dilution

Lanes 1 - 4:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/100000 dilution

Observed band size: 145 kDa

false

Exposure time: 180s